title: Multiplex real-time RT-qPCR for the detection of Norovirus in bottled and tap water using murine norovirus as a process control.
abstract: OBJECTIVE
  To provide a rapid and sensitive method for detecting NoV GI and NoV GII in water and to evaluate the use of the murine norovirus (MNV-1) as a process control.
  
  
  RESULTS
  The method is based on viral concentration by filtration on electropositive filters and direct lysis of adsorbed viruses from filters before RNA extraction and RT-qPCR amplification. An one-step multiplex RT-qPCR assay was developed for the simultaneous detection of NoV GI, NoV GII and MNV-1. Then, water samples were artificially contaminated to determine mean virus recoveries and method sensitivity. The method showed a higher sensitivity for detecting NoV GII (10(3) genome copies per 0·5 l) than for NoV GI (10(4) genome copies per 0·5 l) in the presence of MNV-1 regardless of the type of water. The data also showed that MNV-1 is a robust option as process control.
  
  
  CONCLUSIONS
  The method described provides a valuable tool for the monitoring of potential public health risks associated with NoV contamination in drinkable water.
  
  
  CONCLUSIONS
  Given the increasing evidence for NoV involvement in food outbreaks, the one-step multiplex RT-qPCR assay we used in this study would be a very useful tool to investigate NoV contamination in other food products.
authors.affiliation: Food and Water Virology Unit, ANSES, Maisons-Alfort Laboratory for Food Safety, Maisons-Alfort Cedex, France.
authors.forename: C
authors.initials: C
authors.lastname: Hennechart-Collette
authors.forename: S
authors.initials: S
authors.lastname: Martin-Latil
authors.forename: L
authors.initials: L
authors.lastname: Guillier
authors.forename: S
authors.initials: S
authors.lastname: Perelle
keywords: RT-qPCR; murine norovirus; norovirus; process control; water